Detection of Bacillus cereus group bacteria from cardboard and paper with real-time PCR

Outi Priha (Corresponding Author), Katri Hallamaa, Maria Saarela, Laura Raaska

Research output: Contribution to journalArticleScientificpeer-review

32 Citations (Scopus)

Abstract

The aim of this study was to develop a PCR-based rapid method to detect Bacillus cereus group cells from paper and cardboard. Primers targeting the 16S rDNA and real-time PCR with SYBR green I detection were used in order to be able to also quantify the target. Both autoclaved cardboard samples spiked with B. cereus vegetative cells or spores and naturally contaminated paper and cardboard samples were studied. Results were compared with culturing verified by commercial (API) tests. Several different methods were tested for DNA isolation from the paper and cardboard samples. Two commercial kits intended for soils, the UltraClean soil DNA kit and the FastDNA spin kit for soil, gave the most reproducible results. In spiked samples, the average yield was 50% of added vegetative cells, but spore yield was only about 10%. PCR results from adding vegetative cells correlated with added colony-forming unit (cfu) values (r=0.93, P <0.001) in the range 100–10,000 cfu g−1. Three out of nine studied paper and cardboard samples contained B. cereus group bacteria, based both on culturing and real-time PCR. The numbers were 102–103 bacteria g−1; and PCR gave somewhat higher results than culturing. Thus, real-time PCR can be used as a rapid semi-quantitative method to screen paper and cardboard samples for contamination with B. cereus group bacteria.
Original languageEnglish
Pages (from-to)161 - 169
Number of pages9
JournalJournal of industrial microbiology and biotechnology
Volume31
Issue number4
DOIs
Publication statusPublished - 2004
MoE publication typeA1 Journal article-refereed

Fingerprint

Bacillus cereus
Real-Time Polymerase Chain Reaction
Bacteria
Soil
Spores
Soils
Polymerase Chain Reaction
DNA
Stem Cells
Ribosomal DNA
Application programming interfaces (API)
Contamination

Keywords

  • Bacillus cereus
  • packaging
  • hygiene
  • SYBR green I
  • endospores
  • food

Cite this

Priha, Outi ; Hallamaa, Katri ; Saarela, Maria ; Raaska, Laura. / Detection of Bacillus cereus group bacteria from cardboard and paper with real-time PCR. In: Journal of industrial microbiology and biotechnology. 2004 ; Vol. 31, No. 4. pp. 161 - 169.
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abstract = "The aim of this study was to develop a PCR-based rapid method to detect Bacillus cereus group cells from paper and cardboard. Primers targeting the 16S rDNA and real-time PCR with SYBR green I detection were used in order to be able to also quantify the target. Both autoclaved cardboard samples spiked with B. cereus vegetative cells or spores and naturally contaminated paper and cardboard samples were studied. Results were compared with culturing verified by commercial (API) tests. Several different methods were tested for DNA isolation from the paper and cardboard samples. Two commercial kits intended for soils, the UltraClean soil DNA kit and the FastDNA spin kit for soil, gave the most reproducible results. In spiked samples, the average yield was 50{\%} of added vegetative cells, but spore yield was only about 10{\%}. PCR results from adding vegetative cells correlated with added colony-forming unit (cfu) values (r=0.93, P <0.001) in the range 100–10,000 cfu g−1. Three out of nine studied paper and cardboard samples contained B. cereus group bacteria, based both on culturing and real-time PCR. The numbers were 102–103 bacteria g−1; and PCR gave somewhat higher results than culturing. Thus, real-time PCR can be used as a rapid semi-quantitative method to screen paper and cardboard samples for contamination with B. cereus group bacteria.",
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Detection of Bacillus cereus group bacteria from cardboard and paper with real-time PCR. / Priha, Outi (Corresponding Author); Hallamaa, Katri; Saarela, Maria; Raaska, Laura.

In: Journal of industrial microbiology and biotechnology, Vol. 31, No. 4, 2004, p. 161 - 169.

Research output: Contribution to journalArticleScientificpeer-review

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